Dean G E, Macnab R M, Stader J, Matsumura P, Burks C
J Bacteriol. 1984 Sep;159(3):991-9. doi: 10.1128/jb.159.3.991-999.1984.
The motA and motB gene products of Escherichia coli are integral membrane proteins necessary for flagellar rotation. We determined the DNA sequence of the region containing the motA gene and its promoter. Within this sequence, there is an open reading frame of 885 nucleotides, which with high probability (98% confidence level) meets criteria for a coding sequence. The 295-residue amino acid translation product had a molecular weight of 31,974, in good agreement with the value determined experimentally by gel electrophoresis. The amino acid sequence, which was quite hydrophobic, was subjected to a theoretical analysis designed to predict membrane-spanning alpha-helical segments of integral membrane proteins; four such hydrophobic helices were predicted by this treatment. Additional amphipathic helices may also be present. A remarkable feature of the sequence is the existence of two segments of high uncompensated charge density, one positive and the other negative. Possible organization of the protein in the membrane is discussed. Asymmetry in the amino acid composition of translated DNA sequences was used to distinguish between two possible initiation codons. The use of this method as a criterion for authentication of coding regions is described briefly in an Appendix.
大肠杆菌的motA和motB基因产物是鞭毛旋转所必需的整合膜蛋白。我们测定了包含motA基因及其启动子区域的DNA序列。在该序列中,有一个885个核苷酸的开放阅读框,它极有可能(置信水平为98%)符合编码序列的标准。由295个氨基酸残基组成的翻译产物分子量为31,974,与通过凝胶电泳实验测定的值高度吻合。该氨基酸序列具有很强的疏水性,我们对其进行了理论分析,旨在预测整合膜蛋白的跨膜α螺旋片段;通过这种分析预测出了四个这样的疏水螺旋。可能还存在其他两亲性螺旋。该序列的一个显著特征是存在两段高净电荷密度区域,一段带正电,另一段带负电。文中讨论了该蛋白在膜中的可能组织形式。利用翻译后的DNA序列氨基酸组成的不对称性来区分两个可能的起始密码子。附录中简要描述了将此方法用作编码区域鉴定标准的情况。